Exosome-Depleted FBS

Introducing Gibco Exosome-Depleted FBS

Get the cell culture performance you demand without compromising your results—an ultrapure FBS that provides the highest level of exosome depletion and cell culture performance available. Our exosome-depleted FBS has the highest level of exosome depletion compared to homebrewed material and competitor FBS while retaining the vital nutrients necessary for cell culture. We removed the inefficiency that comes with ultracentrifugation and put the emphasis on a system the removed bovine exosomes while maintaining cell culture performance.

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Hear what Dr. Gilthorpe is saying about Gibco Exosome-Depleted FBS. Saving time, reducing batch-to-batch variation and great growth of cells while getting the highest level bovine exosome depletion are all benefits offered by this product.

“I was able to culture my cells and get very good growth in only 2.5% serum, compared to 10%, due to Gibco Exosome-Depleted FBS, maintaining a lot of the important nutrients ("the good things") that were lost during ultracentrifugation.”Dr. Jonathan Gilthorpe, Pharmacology and Clinical Neuroscience, University, Sweden

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Exosome-Depleted FBS supporting data

72-hour cell growth of rat oligodendrocyte cells with FBS

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Rat oligodendrocyte cells were seeded at 1,000 cells per well in 96-well plates, and grown in medium containing 2% or 10% by volume of one of the following supplements: Gibco Exosome-Depleted FBS, Gibco FBS, the source of the exosome-depleted version), or ultracentrifuged FBS. After 72 hours in culture, the live cells were stained with Invitrogen™ Hoechst 33342, and the plate was imaged and analyzed on a 96-well plate imaging instrument (Trophos Plate RUNNER HD). Results are presented as the total cell count as reported by the 96-well plate analysis. (The results were obtained from the laboratory of Dr. Jonathan Gilthorpe in the department of Pharmacology and Clinical Neuroscience at Umeå University, Sweden.)

Several cell lines were grown in basal medium (DMEM, high glucose, GlutaMAX Supplement) containing 10% Exosome- Depleted FBS or 10% source FBS and assayed for viable cell density (VCD) and cell viability by Vi-CELL instrument analysis. Results are presented as the viability or viable cell density that was achieved in Exosome-Depleted FBS as a percentage of that achieved in the source FBS (used to make the Exosome-Depleted FBS).

Comparison of exosome depletion methods

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Three different lots of FBS were subjected to 3 different methods of exosome depletion. The first used ultracentrigulation at 110,000 X g for 3 hours (labeled as Ultracentrifuge 1). The second method used ultracentrifulation at 150,000 X g for 18 hours (labeled as Ultracentrifuge 2). The third method used our proprietary method for the depletion of exosomes from FBS (labeled as Exosome-Depleted FBS). Source FBS refers to the FBS prior to any exosome depletion process. Afterr the exosome depletions, all samples were subjected to a fluorescence staining assay which involved extracting exosomes using the Total Exosome Isolation Reagent (from serum), and then staining the isolated exosomes with a lipophilic dye. The fluorescent signal is displayed as Relative Fluorescent Units. The data demonstrate the improvement in exosome depletion using our proprieteary method, compared to unltracentrifugation, as evidenced by the much lower fluorescent signals.

The exosome depletion from FBS samples was verified by analysis on a NanoSight instrument (comparing the 30–150 nm count before and after exosome depletion) as well as via fluorescence-based assay. Briefly, this assay involves extracting exosomes from serum using the Total Exosome Isolation Reagent, and then staining the isolated exosomes with Invitrogen BODIPY TR Ceramide. The percent depletion is derived from comparing the fluorescent signal of the exosome-depleted FBS with the source FBS. The first two exosome-depleted lots shown above were produced by our proprietary manufacturing method. Included in the same analysis was a sample of FBS that was ultracentrifuged to deplete exosomes and an exosome-depleted FBS product from a competitor.

72-hour cell growth of rat oligodendrocyte cells with FBS

Click to enlarge

Rat oligodendrocyte cells were seeded at 1,000 cells per well in 96-well plates, and grown in medium containing 2% or 10% by volume of one of the following supplements: Gibco Exosome-Depleted FBS, Gibco FBS, the source of the exosome-depleted version), or ultracentrifuged FBS. After 72 hours in culture, the live cells were stained with Invitrogen™ Hoechst 33342, and the plate was imaged and analyzed on a 96-well plate imaging instrument (Trophos Plate RUNNER HD). Results are presented as the total cell count as reported by the 96-well plate analysis. (The results were obtained from the laboratory of Dr. Jonathan Gilthorpe in the department of Pharmacology and Clinical Neuroscience at Umeå University, Sweden.)

Several cell lines were grown in basal medium (DMEM, high glucose, GlutaMAX Supplement) containing 10% Exosome- Depleted FBS or 10% source FBS and assayed for viable cell density (VCD) and cell viability by Vi-CELL instrument analysis. Results are presented as the viability or viable cell density that was achieved in Exosome-Depleted FBS as a percentage of that achieved in the source FBS (used to make the Exosome-Depleted FBS).

Comparison of exosome depletion methods

Click to enlarge

Three different lots of FBS were subjected to 3 different methods of exosome depletion. The first used ultracentrigulation at 110,000 X g for 3 hours (labeled as Ultracentrifuge 1). The second method used ultracentrifulation at 150,000 X g for 18 hours (labeled as Ultracentrifuge 2). The third method used our proprietary method for the depletion of exosomes from FBS (labeled as Exosome-Depleted FBS). Source FBS refers to the FBS prior to any exosome depletion process. Afterr the exosome depletions, all samples were subjected to a fluorescence staining assay which involved extracting exosomes using the Total Exosome Isolation Reagent (from serum), and then staining the isolated exosomes with a lipophilic dye. The fluorescent signal is displayed as Relative Fluorescent Units. The data demonstrate the improvement in exosome depletion using our proprieteary method, compared to unltracentrifugation, as evidenced by the much lower fluorescent signals.

The exosome depletion from FBS samples was verified by analysis on a NanoSight instrument (comparing the 30–150 nm count before and after exosome depletion) as well as via fluorescence-based assay. Briefly, this assay involves extracting exosomes from serum using the Total Exosome Isolation Reagent, and then staining the isolated exosomes with Invitrogen BODIPY TR Ceramide. The percent depletion is derived from comparing the fluorescent signal of the exosome-depleted FBS with the source FBS. The first two exosome-depleted lots shown above were produced by our proprietary manufacturing method. Included in the same analysis was a sample of FBS that was ultracentrifuged to deplete exosomes and an exosome-depleted FBS product from a competitor.